physicochemical quality control and dosage of total

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Physicochemical quality control and dosage of total polyphenols,flavonoids of Morus alba Leaves (MORACEAE)

Camila B. Pereira*, Aline Marin**, Tiago D. T. Maki**, Raquel M. M.Necchi***, Melânia Palermo Manfron****

ABSTRACT: To establish parameters for quality control of Morus alba leaves were done tests of purityby determining the loss on drying, foreign matter, total ash, acid insoluble ash, swelling index, bitternessindex, yield of crude extract, pH and organoleptic properties. The tests were done with samplescollected on March, June, September and December according to Brazilian Pharmacopeia and WorldHealth Organization. The percentage of loss on drying found are according to the established value of14 %, the highest content was on September 12.89 ± 0.1400 %. The indices of foreign matter found areagreement with the amount allowed by Brazilian Pharmacopeia, which establishes a maximum of 2 %for most of the herbal drugs. The colletc on March showed the highest content of total ash and acidinsoluble ash, respectively, 20.48 ± 0.3517 % and 5.68 ± 0.1057 %. The swelling index showed highervalues in samples collected on September 7.95 ± 0.2186 mL and on December 7.1 ± 0.8544 mL. Thebitterness index was higher in the first two collections, 1.037 units/g in both. The yield produced was 18% on March, 29.61 % on June, 26.47 % on September and 24.35 % on December. The pH of aqueousextract was neutral (pH 7.0) on March and slightly acid (pH 6.0) in other months. The four collectshowed small variations in relation the organoleptic properties. It was done dosage of total polyphenolsand flavonoids with 70% ethanol extracts of M. alba leaves. The content found of polyphenols were6.467 ± 0,012; 5.503 ± 0.007; 6.016 ± 0.020; 6.943 ± 0.046 milligrams of gallic acid per gram of driedplant on March, June, September and December, respectively. The content of flavonoids found were4.345 ± 0.008; 3.782 ± 0.016; 4.362 ± 0.005; 4.893 ± 0.013 milligrams of rutin per gram of dried plant.Descriptors: Morus alba; Moraceae; Quality Control; Leaves.

Controle de qualidade físicoquímico e dosagem de polifenóis totais efavonóides nas folhas de Morus alba (MORACEAE)

RESUMO: Visando estabelecer parâmetros para o controle de qualidade das folhas de Morus albaforam realizados ensaios de pureza através das determinações de umidade, matéria estranha, cinzastotais, cinzas insolúveis em ácido, teor de mucilagem, índice de amargor, rendimento do extrato bruto,propriedades organolépticas e pH. Os ensaios foram realizados com amostras coletadas em março,junho, setembro e dezembro, de acordo com a Farmacopéia Brasileira e a Organização Mundial deSaúde. A perda por dessecação apresentou teor de umidade de acordo com o valor estabelecido de 14%, o valor mais elevado foi em setembro 12.89 ± 0,1400 %. Os índices de matéria estranhaencontrados estão de acordo com a quantidade permitida pela Farmacopéia Brasileira, que estabeleceum máximo de 2% para a maioria das drogas vegetais. A coleta de março apresentou o maior teor de

*Mestranda em Ciência e Tecnologia Farmacêuticas pela Universidade Federal de Santa Maria (UFSM), RS, Brasil.**Aluno de graduação em Farmácia pela Universidade Federal de Santa Maria (UFSM), RS, Brasil.***Mestre em Ciência e Tecnologia Farmacêuticas pela Universidade Federal de Santa Maria (UFSM), RS, Brasil****Doutora em Ciências Biológicas pela Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), SP, Brasil.

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Saúde (Santa Maria), Ahead of Print, v.37,n.2, p. 5768, 2011.

Physicochemicas quality control and dosage of totalpolyphenols, flavonoids of Morus Alba Leaves

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cinzas totais e cinzas insolúveis em ácido, respectivamente, 20,48 ± 0,3517 % e 5,68 ± 0,1057 %. Oíndice de intumescência mostrou valores mais elevados nas amostras coletadas em setembro 7.95 ±0,2186 mL e dezembro 7.1 ± 0,8544 mL. O índice de amargura foi mais elevado nas duas primeirascoletas 1,037 unidades/g em ambas. O rendimento produzido foi 18 % em março, 29,61 % em junho,26,47 % em setembro e 24,35 % em dezembro. O pH do extrato aquoso foi neutro (pH 7,0) em marçoe levemente ácido (pH 6,0) nos outros meses. As quatro coletas apresentaram pequenas variaçõesem relação as propriedades organolépticas. Foi realizada dosagem de polifenóis e flavonóides com oextrato etanólico 70 % das folhas de M. alba. O valor de polifenóis encontrado em março, junho,setembro e dezembro foi 6.467 ± 0,012; 5.503 ± 0.007; 6.016 ± 0.020; 6.943 ± 0.046 miligramas deácido gálico por grama de planta seca, respectivamente. O valor de flavonóides encontrado foi 4.345 ±0.008; 3.782 ± 0.016; 4.362 ± 0.005; 4.893 ± 0.013 miligramas de rutina por grama de planta seca.Descritores: Morus alba; Moraceae; Controle de qualidade; Folhas.

IntroductionThe use of medicinal plants in therapy requires quality of herbal drugs, which begins on

the correct identification of species, planting, harvesting, preparation of plant extracts andmedicines. Therefore, it is necessary to obtain parameters of quality for pharmaceuticalpurposes, which are set out in Pharmacopeias and Officers Codes.

Family Moraceae occurs in tropical and subtropical regions and subdivided intoapproximately 60 genera, which comprise about 1400 species.1 The genus Morus isdistinguished by its inflorescence spike amentacea type. Inflorescence is formed duringmonths of July and August, with small flowers, often singlesex. They have corolla and a cupformed by four free sepals.2 Due to diversity of species of fruits, they are known as whitemulberry (M. alba), blackberry (M. nigra), cranberryred (M. rubra) and bramblerose (M.rosea). The main cultivated are whiteblackberry and blackberry because they don’t havethorns. They are adapted to different soil types, except those with trend water logging. 36 TheMoraceae family leaves are rich in protein, fiber, minerals and vitamin C, they also show atype of glycoprotein, called moran A, which is atributted an antidiabetic effect.7 The genusMorus has a variety of phenolic compounds including isoprenyl flavonoids, coumarins,chromones and xanthones. Many of these compounds exhibit biological properties such asantiinflammatory, diuretic and hypotensive effects.1 The leaves, bark and branch of M. alba(Fig. 1), are used in traditional medicine as antihyperglycemic, to treat fever, to protect theliver, to improve eyesight and to strengthen joints.89 Due to lack M. alba in monographs, thiswork aims to establish standards of quality and purity of plant drug by physicochemicalanalysis and evaluation of total polyphenols and flavonoids in four collections during the year.The physicochemical analysis carried out were: determination of loss on drying, foreignmatter, yield of ethanol extrac, total ash, sulphated ash, swelling index, bitterness index, pHand organoleptic properties.

Saúde (Santa Maria), Ahead of Print, v.37,n.2, p. 5768, 2011.Pereira, C.D., et al.

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Figure 1 — Morus alba leaves.(Font: http://www.cirrusimage.com/tree_white_mulberry.htm)

Material and methods

Plant materialThe Morus alba leaves were collected in March, June, September and December 2010,

in Santa Maria city, Rio Grande do Sul State, Brazil. A sample of plant material was identifiedby Drª Jumaida Maria Rosito and voucher specimens were deposited in the Herbarium ofBiology Department of the Federal University of Santa Maria (UFSM) as voucher specimenSMDB 13079. The plant material was dried at 40 °C with air circulation and grounded in aknife mill.Preparation of ethanol extract

The extracts were obtained by cold maceration of powdered plant drug (80 g) with drugsolvent ratio 20 % (w/v) in 70 % hydroetanolic solvent during 30 days, after this, they wereconcentrated, lyophilizated and calculated yield.

Physicalchemistry quality controlLoss on drying

The water determination was performed according to Brazilian Pharmacopeia.10 It wasweighed exactly 1 g of powdered plant drug and it underwent heating in an oven at 105 °Cduring 5 hours. The loss of drying weight was calculated considering the airdried sample.The analysis was performed in triplicate.



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Foreign matterMacro e microscopy examination was conveniently employed to determine the presence

of foreign matter in powdered plant drug according to Brazilian Pharmacopeia.10 In themacroscopy examination, 25 g of plant drug was spread on a thin layer from which the foreignmatter was selected by inspection through the magnifying less. The content of foreign matterwas calculated considering the airdried sample. The analysis was performed in triplicate.Total ash

The analysis of total ash was conducted according to Brazilian Pharmacopeia.10 Thepowdered plant drug (3g) was accurately weighed into a crucible that has been previouslyignited, cooled and weighed. Then the sample was ignited by gradual heating to 450 °C,cooled in a desiccator and weighed. For obtaining carbonfree ash, the residue was moistenedwhit 2 mL of water after cooling the crucible. The remaining material was dried on a waterbathand then placed on a hotplate, being ignited again until constant weight. The residue wascooled in a desiccator for 30 minutes and weighed. Total ash content was calculatedconsidering the dry material. The analysis was performed in triplicate.Acid insoluble ash

The analysis of acid insoluble ash was conducted according to Brazilian Pharmacopeia.10In the crucible containing the total ash, it was added 25 mL of hydrochloric acid (70 g/L),covered with a watchglass and boiled gently for 5 minutes. The watchglass was rinsed with 5mL of hot water and then this liquid was added to the crucible. The insoluble matter wascollected on an ashless filterpaper and washed with hot water until the filtrate was neutral.Thereafter, the filterpaper containing the insoluble matter was transferred to the originalcrucible, which was dried on a hotplate and ignited to constant weight. After cooling theresidue in a desiccator, the crucible was weighted and the content of acidinsoluble ashcalculated considering the dry material. The analysis was performed in triplicate.Swelling index

The analysis of swelling index was conducted according to Brazilian Pharmacopeia.10One gram of powdered plant drug was introduced into a 25 mL glassstoppered measuringcylinder for the verification of the volume occupied by the sample. Next, 25 mL of water wasadded and the mixture was shaken thoroughly every 10 minutes for about 1 hour. After that,the mixture was stood for 3 hours at room temperature, and finally the volume in ml occupiedby the plant material was remeasured. The test was performed in triplicate.Bitterness index

The determination of the index of bitterness was performed according to BrazilianPharmacopeia.10 The bitter properties of plant material were determined by comparing the


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threshold bitter concentration of the extract from the material with the concentration from adilute solution of quinine hydrochloride. Drinkingwater was the vehicle to the extraction ofplant materials and for the mouthwash after each tasting. It’s was prepared a stock solution(St) of M. alba at 1 % (w/v). St was transferred to 10 tubes with volumes from 1 to 10 mL. Thestandard solution of quinine hydrochloride R was distributed in nine tubes from 0.042 to 0.058mg/10 mL. The birtteness value was expressed in units equivalent to the bitterness of asolution containing 1 g of quinine hydrochloride in 2000 mL.Yield

The analysis of yield was conducted according to Costa.11 The yield was determinedusing the equation: Yield (%) = Mf / Mi X 100, where Mi is the initial mass of sample (g) and Mfis the final mass of the dry extract (g). Crude extracts were obtained by cold maceration ofpowdered plant drug (80 g) with drug solvent ratio 20% (w/v) in 70 % hydroetanolic solventduring 30 days and the yield determined. Botanical studies of yield must be performed as anindicative of correct amount of material to be collected for production of extracts and fractionsused in pharmacological tests.Organoleptic properties and pH

The organoleptic features analyzed were odour, flavor and color. The odour was checkedby pressing of a small portion of leaves on the palm of the hand, inhaling slowly andrepeatedly. The color of the leaves was analyzed with fresh plant under diffuse daylight. Theflavor was tasted through the instillation of few drops on tongue of an aqueous solution 1 %(w/v) of the powdered plant drug heated on an eletricplate and brought to be boiling for 5minutes.12 pH was verified by potentiometry with the above solution, according to BrazilianPharmacopeia.13

Dosage of total polyphenolsThe dosage of polyphenols was performed through the FolinCiocalteau, with some

modifications.1415 Based on the solution of 0.04 % of the ethanol extract of M. alba, solutionswas prepared in volumetric flasks at concentrations of 5,10,20,30 µg/mL. To this end, we usedthe required amount of solution of 0.04 % with addition of 1 mL of FolinCiocalteau for 5minutes, 2 mL of sodium carbonate (Na2CO3) 20 % for 10 minutes. The solutions werehomogenized, capped and protected from light and kept at room temperature. Theabsorbance was measured at 730 nm using water as a blank. The test was performed intriplicate. The standard curve was obtained using standard solutions of gallic acid at the sameconcentration of the sample. The content of total polyphenols was expressed as milligram ofgallic acid equivalent per gram of dried plant.



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Dosage of flavonoidsThe dosage of flavonoids was performed according to a modified methodology from

Rio16. Based on the solution of 0.04 % of the ethanol extract of M. alba, solutions wasprepared in volumetric flasks at concentrations of 10,20,50,100,150 µg/mL . To this end, weused the required amount of solution of 0.04 % with addition of 75 l of 5 % methanol(MeOH) solution of AlCl3, 3.9 mL of methanol 70 % for 30 minutes. The solutions werehomogenized, capped and protected from light and kept at room temperature. The absorbancewas measured at 425 nm using MeOH 70% as a blank. The test was performed in triplicate.The standard curve was obtained using standard solutions of rutin at the same concentrationof the sample. The flavonoids content was expressed as milligram of rutin equivalent per gramof dried plant.Statistical analysis

Results were expressed as mean ± standard error of mean (SEM). Statistical analysiswas performed using Oneway ANOVA followed by Tukey test using Graph Prism 5.0 software(Graph Pad Software, San Diego, CA, USA) and the results were considered significant whenp < 0.05.

Results and discussionPhysicoChemical Quality Control

The loss on drying of M. alba leaves showed a moisture content according to theestablished value of 14 %, which is the maximum limit recommended of moisture for plantdrugs.12;13;17 According to Simões18 the water founded in the plant drug is directly related to itsstorage and microbial contamination or enzymatic degradation by action of the chemicalconstituents. The excess of water in the raw material enables the development of fungi andbacteria.19 Therefore, preliminary operations employed in the leaves were effective instandardizing of moisture content to established values. Through statistical analysis werechecked significant differences in the loss on drying in the four collections (table 1).

Table 1 — Loss on drying, mean ± standard error of mean (SEM) of Morus alba leaves.

Indices of foreign matter found in M. alba on March and June are agreement with theamount allowed by Brazilian Pharmacopeia,10 which establishes a maximum of 2 % for most ofthe herbal drugs (table 2). Collections made during September and December didn’t show

Month Mean (%) SEMMarch 9.73 ± 0.1253June 11.15 ± 0.2118

September 12.89 ± 0.1400December 8.56 ± 0.0808


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foreign matter. There weren’t significant differences in the indices of foreign matter in the fourcollections. The amount of foreign matter may be related to the fact of leaves have been preselected and milled.20 Analysis for determination of foreign matter has the purpose to identifyportions or products not specified in the definition and description of the drug, besidesimpurities from mineral or organic nature, don’t related to the drug.13

Table 2 — Foreign matter, mean ± standard error of mean (SEM) of Morus alba leaves

The determination of contenting total ash is important for quality control, since its aim is tocheck the presence of nonvolatile inorganic impurities that may contaminate the plantdrug.10;21 The colletc that had the highest content of total ash was done on March (table 3).This can be explained by the higher amount of inorganic salts present in the first collection.17

In the analysis of acid insoluble ash was possible to observe a large discrepancy invalues. In the gathering done on March and December, the content found was 5.68 % and3.50 %, respectively. Through statistical analysis were checked significant differences in thecontent of total ash and acid insoluble ash in the four collections. Determination of ashinsoluble in acid chloridric is intended to detect constituents of silica and siliceousconstituients, wich above of the established for the plant drug indicates contamination byexcess soil or sand.13 There is not maximum value for total ash and acid insoluble ash officiallyestablished for the species in study.

Table 3 — Determination of total ash, acid insoluble ash, mean ± standard error of mean (SEM) of Morus alba leaves.

Analyzing the swelling index, M. alba leaves showed the highest values in samplescollected on September and December (Table 4). The significant differences were checkedonly in results obtained on September versus March and September versus June. Banderó 22found a quantity of swelling index of 2.03 mL to Glechon spathulata. This test serves tomeasure the volume (mL) occupied by plant matter plus mucilage or other materialattached.12,13,17

Month Mean (%) SEMMarch 0.0200 ± 0.0121June 0.0126 ± 0.0063

Month Total ash (%) Acid insoluble ash (%)March 20.48 ± 0.3517 5.68 ± 0.1057June 9.57 ± 0.4908 0.75 ± 0.0333

September 11.88 ± 0.2408 1.67 ± 0.0738December 17.86 ± 0.4020 3.508 ± 0.0601



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Table 4 — Swelling index, mean ± standard error of mean (SEM) of Morus alba leaves.

Pharmacopeias and specialist books on herbal drugs include certain types of sensationlike flavour. Analysis of the bitterness index is determined by comparing the thresholdconcentration of bitterness of an extract and the dilute solution of quinine chloride. Thus, thereare astringent, oily, mucilaginous, bitter and pungent tastes. The bitterness index was higher inthe first two collections (table 5), it shows that the drug contains bitter substances, especiallyconsidering the dilution used in the test. To Wasicky et al.,23 bitterness index has solid basestheoretical and experimental, which competes with the best biological methods. However,Mello et al., 24 affirmed that this test may introduce errors due to individual variability, since itdoesn’t statistically reproducing real information. Thus, to perform this test it is necessary aprevious training of experimenter, which combined with statistical analysis, allow that thebiological variability be minimized, becoming a reliable method.

There were differences in about 11 % in yield’s assessment of 70 % ethanol extracts. Thehighest yields were on June and September. The pH of the aqueous extract was neutral (pH7.0) on March and slightly acid (pH 6.0) in other months, this suggests presence of neutral andacid substances, respectively, in M. alba leaves.

Table 5 — Bitterness index, yield, pH of Morus alba leaves.

In relation the organoleptic properties, the four collect showed small variations, they wereodorless and the aquous extract showed a slight bitter flavor. The fresh leaves showed a lightgreen color in the collection of March, midgreen on June and September and dark green onDecember. Total polyphenols content found in ethanol extracts of M. alba on March, June,September and December were 6.467 ± 0,012; 5.503 ± 0.007; 6.016 ± 0.020; 6.943 ± 0.046milligrams of gallic acid per gram of dried plant (figure 2). The regression curve obtained was Y= 0.2879x – 0.0685 (R = 0.9912). The highest content of polyphenols was found in collectionon December (6.943 ± 0.046 mg/g). The content of flavonoids found was 4.345 ± 0.008; 3.782± 0.016; 4.362 ± 0.005; 4.893 ± 0.013 milligrams of rutin per gram of dried plant. Theregression curve obtained was Y= 0.1821x – 0.1545 (R = 0.9935).

Month Mean (mL) SEMMarch 5.83 ± 0.1202June 5.73 ± 0.0882

September 7.95 ± 0.2186December 7.1 ± 0.8544

Month Bitterness index (units/g) Yield (%) pHMarch 1.037 18.0 7June 1.037 29.61 6

September 0.84 26.47 6December 0.84 24.35 6


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Figure 2 — Contents of polyphenols and flavonoids in ethanol extracts produced in four seasons.

Plants can accumulate phenolic compounds in their metabolism under various stressconditions.25,26 Second Paliyath et al.,27, most plants suffer physiological and biochemicaldamage by exposure to different temperatures. Sývacý et.al., 28 found significant differences inpolyphenol content of M. nigra and M. alba stem in the four seasons.

Through statistical analysis used in this study, was checked significant differences in thepolyphenol and flavonoids content in four collections, only on March versus September therewere not significant differences in flavonoids content. This fact can be explained by changes inclimate, collection time, soil formation, plant age, temperature and total rainfall. The informationobtained in this study has been relevant to characterization and physico chemical qualitycontrol of M. alba leaves.

ConclusionDue to widespread use of M. alba leaves in traditional medicine, it is necessary to obtain

quality standard for the use of this plant. The methods employed were adequate to assess thequality control of vegetable drugs in study, since it hasn’t description in Pharmacopoeias. Thedosages of polyphenols and flavonoids in the four colletions showed higher content in summer.

References

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5. Donadio LC. Dicionário das frutas. Jaboticabal. 2007; p. 300.6. Gomes P. Fruticultura brasileira 13ª ed. São Paulo: Nobel. 2007; p. 446.7. Andallu B, Vardacharyulun CH. Effect of mulberry leaves on diabetes. Int. J. Diab. Dev. Countries. 2001;21: 14751.8. Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and theirscavenging effects in superoxide radicals. Food Chemistry. 1999; 64: 555–59.9. Kim SU, Kim LHS, Kim I, MY, Kim, KS Ahn. Determination of 1deoxynojirimycin in Morus alba L. leavesthrough derivatization with 9fluorenylmethyl chloroformate followed by reversedphase high performanceliquid chromatography. Journal of Chromatography. 2003; 1002:93–9.10. Farmacopéia Brasileira. 4. ed. São Paulo: Atheneu. 200011. Costa AF. Farmacognosia. Lisboa: Fundação Calouste Gulbenkian. 4 ed.; 1994; v.2, p. 1117 .12. World Health Organization. Quality Control Methods for Medicinal Plant Materials. Geneva, 1998, p.122.13. Farmacopéia Brasileira. 4. ed. São Paulo: Atheneu. 1988.14. Simonovska B, Vovk I, Andrensek S, Valentova K, Ulrichova J. Investigation of phenolic acids in yacon(Smallanthus sonchifolius) leaves and tubers. Journal of Chromatography. 2003; 1016(1), 8998.15. Kumazawa S, Hamasaka T, Nakayama T. Antioxidant activity of propolis of various geographic origins.Food Chem. 2004; 84:32939.16. Rio RGW. Métodos de controle químico de amostras de própolis. Dissertação de mestrado. Universidadede São Paulo, São Paulo. 199617. Hänsel R, Sticher O, Steinegge RE. Pharmakognosie – Phytopharmazie.Berlin: Springer. 199918. Simões CMO, Mentz LA, Schenkel EP, Irgang BE, Stehmann JR. Plantas da medicina popular no RioGrande do Sul. 5. ed. Porto Alegre: Ed. Universidade/UFRGS. 1998, p. 173.19. Farias, MR. Avaliação da qualidade de matériasprimas vegetais In: Simões, C.M.O. et al (Org.).“Farmacognosia: da planta ao medicamento”. 5. ed. Porto Alegre/Florianópolis: Ed. Universidade UFRGS/Ed. da UFSC. 200320. Frasson, APZ, Bittencourt CF, Heinzmann BM. Caracterização físico química e biológica do caule deCaesalpinia ferrea Mart. Rev. Bras. Farmacog. 2003; 13: 359.21. Sonaglio D, Ortega GG, Petrovick PR, Bassani VL. Desenvolvimento tecnológico e produção efitoterápicos. In: Simões C.M.O, E.P. Schenkel, M.J.C.P. Gosmann, L.A. Mentz., P.R. Petrovick.Farmacognosia: da planta ao medicamento. 5.ed. Porto Alegre/Florianópolis: Editora da UFRGS/ Editora daUFSC. 200322. Banderó VC. Physicochemical characterization for qualiy control of Glechon spathulata. Dissertação deMestrado, Progama de Pósgraduação em Ciências Farmacêuticas, Universidade Federal de Santa Maria.200923. Wasicky R, Barbieri E, Weber H. Contribuição para o método de dosagem de princípios ativos em drogase preparações pelo amargor. Anais da Faculdade de Farmácia e Odontologia da Universidade de São Paulo.1943; 3: 11319.24. Mello JCP, Petrovick PR. Quality control of Baccahris trimera (Less) DC (Asteraceae) hydroalcoholicextracts. Acta Farm Bonaerense. 2000; 19: 21115.25. Pasqualini V., Robles C., Garzino S., Greff S., Bousquet MA, Bonin G.. Phenolic compounds content inPinus halepensis Mill needles: a bioindicator of air pollution. Chemosphere. 2003; 52:23948.26. Christie PJ, Alfenito MR, Walbot V. Impact of lowtemperature stress on general phenilpropanoid andanthocyanin pathways: enhancement of transcript abundance and anthocyanin pigmentation in maize


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seedlings. Planta. 1994; 194: 54149.27. Paliyath G., Pinhero RG, Rao MV, Murr DP. Fletcher RA. Changes in activities of antioxidant enzymesand their relationship to the genetic and paclobutrazolinduced chilling tolerance in maize seddlings. PlantPhysiol. 1997; 114:695704.28. Sývacý A., Sökmen M. Seasonal changes in antioxidant activity, total phenolic and anthocyaninconstituent of the stems of two Morus species (Morus alba L. and Morus nigra L.) Plant Growth Regulation.2004; 44: 3251256.

Camila Bugnotto PereiraEndereço para correspondência — Programa de PósGraduação em Ciências Farmacêuticas,UniversidadeFederal de Santa Maria, Prédio 26, Santa Maria, RS, Brasil.CEP: 97105900Email: [email protected]ículo lattes: http://lattes.cnpq.br/5954851418582018

Recebido em 01 de junho de 2011.Aprovado em 30 de agosto de 2011.



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physicochemical quality control and dosage of total

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